Role of O-linked carbohydrate of human urinary trypsin inhibitor on its lysosomal membrane-stabilizing property.
Human urinary trypsin inhibitor (UTI) was digested with various enzymes to obtain O-glycoside linked N-terminal glycopeptide (UTIm1), N-glycoside linked C-terminal tandem Kunitz-domains (domain I and II, UTIm2), UTI lacking O-glycoside (UTIc), asialo UTI (UTIa) and UTI lacking N-glycoside (UTIn). We investigated the membrane stabilizing effect of these UTI derivatives on rat renal lysosome by measurement of lysosomal enzyme N-acetyl-beta-D-glucosaminidase (NAG) release after hypotonic treatment. Intact UTI suppressed NAG release, but aprotinin, gabexate mesilate (FOY), nafamostat mesilate (FUT) and recombinant domain II of UTI (R-020) had no effect, indicating that inhibition of serine proteases was not involved and the carbohydrate moiety of UTI might be necessary for this property. Among UTI derivatives, UTIm1, UTIm2, UTIm1+ UTIm2, and UTIc had no effect. In contrast, UTIa or UTIn suppressed NAG release. From these results, we conclude that O-glycoside linked core protein without N-glycoside is essential to the lysosomal membrane-stabilizing property of UTI.
Kato
Y
Y
Fuji Central Research Laboratory, Mochida Pharmaceutical Co., Ltd., Shizuoka, Japan.
Kudo
M
M
Shinkawa
T
T
Mochizuki
H
H
Isaji
M
M
Shiromizu
I
I
Hoshida
K
K
D000118:Acetylglucosaminidase / Q000378:metabolism; D000818:Animals; D002462:Cell Membrane / Q000378:metabolism*; D006020:Glycopeptides / Q000494:pharmacology; D006023:Glycoproteins / Q000737:chemistry / Q000494:pharmacology*; D006027:Glycosides / Q000494:pharmacology; D006801:Humans; D007668:Kidney / Q000737:chemistry; D008247:Lysosomes / Q000378:metabolism*; D008562:Membrane Glycoproteins / Q000502:physiology*; D008666:Metalloendopeptidases / Q000378:metabolism; D009005:Monosaccharides / Q000032:analysis; D009844:Oligosaccharides / Q000737:chemistry / Q000494:pharmacology*; D051381:Rats; D011994:Recombinant Proteins / Q000494:pharmacology; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology
D006020:Glycopeptides; D006023:Glycoproteins; D006027:Glycosides; D008562:Membrane Glycoproteins; D009005:Monosaccharides; D009844:Oligosaccharides; D011994:Recombinant Proteins; D015842:Serine Proteinase Inhibitors; D000118:Acetylglucosaminidase; D008666:Metalloendopeptidases; C028665:urinastatin